Efficient transgene-free multiplexed genome editing via viral delivery of an engineered TnpB.

Trevor J. Weiss; Maris Kamalu; Honglue Shi; Gabriel Wirnowski; Alice Ingelsson; Jasmine Amerasekera; Kamakshi Vohra; Marena I. Trinidad; Z. M. Li; Emily dos Santos Galante Freitas; Noah Steinmetz; Charlie Ambrose; Kerry Chen; Jennifer A. Doudna; Steven E. Jacobsen

doi:10.1111/pbi.70644

Plant BiologyPlant Biotechnology JournalMar 17, 2026

Peer-reviewed journal article

Efficient transgene-free multiplexed genome editing via viral delivery of an engineered TnpB.

Trevor J. Weiss iD, Maris Kamalu iD, Honglue Shi iD, Gabriel Wirnowski iD, Alice Ingelsson, Jasmine Amerasekera, Kamakshi Vohra iD, Marena I. Trinidad iD, Z. M. Li, Emily dos Santos Galante Freitas, Noah Steinmetz, Charlie Ambrose iD, Kerry Chen, Jennifer A. Doudna iD, Steven E. Jacobsen iD

Abstract

Virus-induced genome editing (VIGE) using compact RNA-guided endonucleases is a transformational new approach in plant biotechnology, enabling tissue-culture-independent and transgene-free genome editing (Hu et al. 2025; Liu et al. 2025; Weiss et al. 2025). We recently established a VIGE approach for heritable editing at single loci in Arabidopsis by delivering the compact genome editor ISYmu1 TnpB (Ymu1) and its guide RNA (gRNA) via Tobacco Rattle Virus (TRV) (Weiss et al. 2025). Here, we greatly improved this approach by devising a multiple gRNA expression system and by utilizing an engineered high-activity Ymu1 variant (Ymu1-WFR) (Zhou et al. 2026) to develop an efficient multiplexed genome editing platform.

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